US9696536B2

US9696536B2 - Arrangement for changing the focus of a microscope objective

Info

Publication number: US9696536B2
Application number: US15/009,195
Authority: US
Inventors: Thomas Nobis
Original assignee: Carl Zeiss Microscopy GmbH
Current assignee: Carl Zeiss Microscopy GmbH
Priority date: 2015-01-28
Filing date: 2016-01-28
Publication date: 2017-07-04
Anticipated expiration: 2036-01-28
Also published as: US20160216499A1; DE102015201393A1

Abstract

A plan apochromat corrected microscope objective including multiple subsystems with optical components and/or component groups, wherein an optical component or a component group can move axially in the interior of the microscope objective. The axially movable component or the component group includes a concave-convex lens or lens group oriented toward the object plane, with a form factor of:

X = \frac{c_{1} + c_{2}}{c_{1} - c_{2}}

wherein X is the form factor, c₁is the curvature of the surface oriented in the direction of the object plane, and c₂is the curvature of the surface oriented in the direction of the image plane, of the axially movable lens or lens group, X lying within a range from −8<X<−1.

Description

RELATED APPLICATIONS

The present application claims priority to German Application No. 102015201393.6, filed Jan. 28, 2015, said application being incorporated herein in its entirety by reference.

TECHNICAL FIELD

The invention relates to an arrangement for changing the focus of a microscope objective, particularly a plan apochromat corrected microscope objective which is composed of multiple subsystems comprising optical components and/or component groups, wherein at least one optical component or one component group can also move axially in the interior of the microscope objective.

BACKGROUND

In modern, wide-field and/or laser-scanning microscopy with biological or other samples, it is essential that optical planes at different sample depths (z stacks) are quickly captured at a high numerical aperture (NA>0.8 with water immersion).

There are various different solutions for changing the focus in the known prior art. One of the solutions is the movement of the entire microscope objective. This method transmits vibrations to the sample if an immersion medium is used, such as water or oil, for example. In addition, the speed which can be achieved is limited by the high mass of the microscope objective.

The use of adaptive elements for changing the focus is also known. In this case, it is disadvantageous that it is necessary to insert an additional optical arrangement into the beam path to constitute the pupil. This is also very unwieldy.

Moreover, it is not possible to correct field aberrations in wide-field microscopy, such that the focal range of this solution is very restricted.

Another known solution is a movement of lenses in the tube optics used. Because of the low numerical aperture in the tube optics and the resulting low depth imaging factor, long movement paths are necessary in this configuration. In addition, aberrations arising on the lenses close to the object due to defocus are difficult or sometimes even impossible to correct at high numerical apertures.

In addition, a solution is known in which “Alvarez elements” are used for changing the focus. These are specially ground plates which generate a focal shift when moved toward each other. A dedicated access to the pupil is also necessary in this case. This positioning can be achieved in the objective due to the Alvarez plates functioning in the transmission path. However, this increases the technical complexity of the microscope objective significantly.

The typical approach for photographic objectives of moving lenses axially in the inside of the objective has led in the past to relatively little success. Due to the high numerical aperture, as well as the wide angle of incident light rays at the edge of the objective, strong aberrations are typically produced—in addition to a focal shift—when the lenses are moved. To correct these, a movement of additional lenses is necessary.

For this reason, in typical correction objectives with a long movement path, multiple lenses are typically moved. However, these coupled movements cannot be carried out very quickly.

By way of example, DE 102005034441A1 describes a microscope objective having an optical frontal element, multiple optical elements which are spaced apart from the frontal element and from each other, and an adjusting device, wherein at least one of the optical elements can be moved along the optical axis by means of the adjusting device in such a manner that the position of the focus of the objective is displaced relative to the frontal element in the direction of the optical axis and/or an aberration of the objective caused by temperature is compensated.

In this solution, the action of focusing at a high numerical aperture is carried out with movable elements. In this case, a diffractive element is moved. A substantial disadvantage in this case is that light is scattered by diffraction at other orders than the functional order.

Proceeding from the solutions according to the prior art, the problem addressed by the invention is that of further advancing an arrangement for focusing a microscope objective, in such a manner that it is possible to quickly change the focus, using simple means, over a relatively large focal range while avoiding vibrations and aberrations.

SUMMARY

This problem is addressed by an arrangement of the type described above.

According to embodiments of the invention, the axially movable component or the component group consists of a concave-convex (crescent-shaped) lens or lens group oriented toward the object plane, with a form factor of:

X = \frac{c_{1} + c_{2}}{c_{1} - c_{2}}

wherein X is the form factor, c₁is the curvature of the surface oriented in the direction of the object plane, and c₂is the curvature of the surface oriented in the direction of the image plane, of the axially movable lens or lens group, and wherein X lies within a range from −8<X<−1.

Because the focus change is carried out in the interior of the microscope objective, vibrations are avoided. Due to the relatively low mass of a single lens or lens group, it is possible to quickly change the focus with a comparatively large focal shift (more than 100 focal depths).

In this configuration, the image quality, as regards axial aberration, spherical aberration, and field aberration, is preserved.

The specially shaped, axially movable single lens or lens group in this case is arranged, proceeding from the object plane, behind a first optical component or a component group, called the frontal group. The total focal length f_frontof the frontal group, relative to the focal length of the objective f_objectivelies in the range of:
f _front<−4×f _objectiveor f _front>4×f _objective
and the focal length of the movable single lens or lens group, for an axial range of travel between approximately 0 mm and 5 mm, is within the range of:
3×f _objective <f _ShiftLens<20×f _objective.

Both the frontal group and the component which is arranged with axial movement for the purpose of changing the focus can in this case either consist of a single lens, a double lens component, or a triple lens component. Combinations of two of these elements can also be contemplated.

If the focal length is made too large, the sensitivity for the change of focus is lowered, which means that the lens or lens group must be moved unreasonably far to ensure a change of focus in the sample. If, in contrast, the focal length is made too small, the lens movement generally produces distortions.

Within the ranges named above, depending on the type of objective, the form factor and focal length (two degrees of freedom) are selected such that no spherical aberration occurs on the axis and no coma occurs on the edge of the field during the movement (two requirements). For the selected lens, there is usually an existing solution at multiple positions in the objective to fulfill both requirements in the two degrees of freedom available.

Due to the position of the arrangement of the axially movable single lens or lens group, the astigmatism on the edge of the field is minimized over the focal range. The specially designed concave-convex shape (crescent shape) is also beneficial for the imaging properties of the objective when the single lens or lens group is unadvanced, because it has an essentially aplanatic shape.

In addition, the selection of the moved single lens or lens group in the frontal region of the objective leads to an early compensation of the beam path changed by the change of focus. Light passes through the rear portion of the objective in a similar manner for all focus positions. The result is fewer induced aberrations.

A further effect when the arrangement according to the invention is used is a special color correction. If a single lens is used as the movable component, the Abbe coefficient/dispersion of the material is selected according to the invention in such a manner that no axial aberration occurs during the movement, in cooperation with the color properties of the frontal and rear groups. Because this can happen regardless of the selection of the refractive index of the glass for the primary wavelengths, the properties of the correction mentioned above are not changed. If no glass is available for this purpose, the relevant Abbe coefficient can be produced via a lens component.

The arrangement according to the invention for the change of focus can be used with positive results in, by way of example, 25×/0.8 W, 25×/0.9 W, and 20×/1.0 W microscope objectives, although other designs can be contemplated.

In this case, the arrangement can be integrated in already existing microscope objectives with a fixed focal length, with minimal modifications.

Furthermore, the arrangement according to the invention is elevated above the prior art to the following extent:

The two air spaces before and after the axially movable single lens or lens group are generally used in a fixed focus objective for adjustment procedures. This means that spherical aberration on the axis is adjusted one time, wherein both air spaces are lengthened simultaneously. By moving the single lens or lens group—that is, by reducing one of the two air spaces and enlarging the other air space, the axial aberration on the axis is typically adjusted one time.

This adjustment is intentionally modified in the arrangement according to the invention. A correction mechanism is included which enables a cover glass thickness correction and therefore can also be used for adjusting the spherical aberration. The axial aberration, in contrast, can be adjusted simply by changing the distance between the frontal and rear group, as a result of the correction named above. The air spaces before and after the axially movable single lens or lens group are specifically enlarged. Objectives in the prior art use very closely spaced lenses in the frontal objective part, which contribute to the rapid reduction in the angle of incidence of light on the lens surfaces, as well as to the reduction in the objective diameter. This was specifically circumvented in the arrangement according to the invention in order to create more space for the lens movement.

The above summary is not intended to describe each illustrated embodiment or every implementation of the subject matter hereof. The figures and the detailed description that follow more particularly exemplify various embodiments.

BRIEF DESCRIPTION OF THE DRAWINGS

Subject matter hereof may be more completely understood in consideration of the following detailed description of various embodiments in connection with the accompanying figures, in which:

FIG. 1 shows a schematic illustration of the single lens of the arrangement according to the invention, with the curvature radii in a plan-apochromat 25×/0.9 W for a focal range of 200 μm;

FIG. 2 shows a schematic illustration of the thicknesses of the single lenses in FIG. 1, as well as the air spaces;

FIG. 3 shows schematic illustrations of the arrangement according to the invention in FIG. 1, for various configurations;

FIG. 4 shows schematic illustrations of transverse aberrations of the arrangement in FIG. 1;

FIG. 5 shows a schematic illustration of the single lenses of the arrangement according to the invention, with the curvature radii in a plan apochromat 25×/0.8 W for a focal range of 250 μm in an objective modification known from U.S. Pat. No. 7,782,539;

FIG. 6 shows a schematic illustration of the thicknesses of the single lenses according to FIG. 5, as well as the air spaces;

FIG. 7 shows schematic illustrations of the arrangement according to the invention in FIG. 5, for various focal depths in the sample;

FIG. 8 shows schematic illustrations of transverse aberrations of the arrangement in FIG. 5;

FIG. 9 shows a schematic illustration of the single lenses of the arrangement according to the invention, with the curvature radii in a plan apochromat 20×/1.0 W for a focal range of 300 μm in an objective modification known from DE 102011109783;

FIG. 10 shows a schematic illustration of the thicknesses of the single lenses in FIG. 9, as well as the air spaces;

FIG. 11 shows schematic illustrations of the arrangement according to the invention in FIG. 9, for various focal depths in the sample; and

FIG. 12 shows schematic illustrations of transverse aberrations of the arrangement in FIG. 9.

While various embodiments are amenable to various modifications and alternative forms, specifics thereof have been shown by way of example in the drawings and will be described in detail. It should be understood, however, that the intention is not to limit the claimed inventions to the particular embodiments described. On the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the subject matter as defined by the claims.

DETAILED DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the arrangement according to the invention in a plan apochromat 25×/0.9 W for a focal range of 200 μm.

Proceeding from the object plane OE, the arrangement consists of a frontal group FG, having lenses L1 and L2, an axially movable lens L3 designed as the focus lens FL, and a rear group RG with the lenses L4 to L12. FIG. 1 also shows the radii r1 to r18 of the individual optical components.

The schematic illustration of the arrangement according to the invention in FIG. 2 complements the illustration in FIG. 1 by adding the thicknesses D1 to D12 of the lenses 11 to [L12], and the air spaces LA1 to LA6.

Because of the axial movement of the concave-convex (crescent-shaped) lens 2—that is, of the focus lens [FL]—in combination with the form factor X thereof, which is:

X = \frac{c_{1} + c_{2}}{c_{1} - c_{2}}

wherein c₁is the curvature of the surface oriented in the direction of the object plane, and c₂is the curvature of the surface oriented in the direction of the image plane, of the axially movable lens or lens group, and wherein X lies within a range from −8<X<−1, the plan apochromat lens can be focused quickly with no vibration.

For a focal range of 200 μm, a numerical aperture of 0.9, a magnification factor β with a 25× larger image, and when an immersion agent/sodium chloride solution W is used, with a field number of 18, this embodiment is characterized by the following design data, wherein r1 to r18 are the curvature radii of the optical components in mm, D is the thickness of the air spaces between the optical components in mm, LA1 to LA6 are the air spaces between the optical components in mm, n_eis the refractive index, and v_eis the Abbe coefficient:


	Curvature	Size		Abbe
	radius	Air space	Refractive	Co-
	R	D, LA	Index	efficient
Lens No.	[mm]	[mm]	n_e	v_e

Sample, cell medium	flat	P	1.38	63.0
Cover glass	flat	DG	1.53	54.3
Immersion	flat	AA	1.34	55.4

1

flat

D1

0.50

1.49

70.2

2

r1

−1.636

D2

5.79

1.89

40.5

r2

−6.167

LA1

3

r3

−14.549

D3

2.40

1.59

61.0

r4

−9.422

LA2

4

r5

70.312

D4

3.00

1.81

46.2

r6

−22.736

LA3

5	r7	42.712	D5	3.80	1.49	84.1
6	r8	−18.905	D6	1.60	1.66	39.5
7	r9	13.868	D7	3.90	1.50	81.1

r10

−18.518

LA4

8	r11	−344.203	D8	1.40	1.64	42.2
9	r12	8.092	D9	5.04	1.53	76.6
	r13	−48.460	LA5	0.35
10	r14	8.015	D10	3.80	1.59	61.0
11	r15	11.802	D11	1.40	1.72	53.6
	r16	5.902	LA6	5.14
12	r17	−6.704	D12	4.69	1.81	46.2

r18	−9.177	0.13
		126.50

and a tube lens arranged behind the same, with a focal length of f=164 mm.

Depending on the sample thickness P, the cover glass DE which is used, and the working distances AA, the following values are then found for the air spaces LA1 to LA4. The air spaces LA5 and LA6 are always the same in this case.


P	DG	AA	LA1	LA2	LA3	LA4
[mm]	[mm]	[mm]	[mm]	[mm]	[mm]	[mm]

0	0	0.56	1.78	0.49	2.47	0.3
0.2	0	0.56	0.95	1.33	2.47	0.3
0	0.17	0.4	1.78	0.49	1.93	0.84
0.2	0.17	0.4	0.95	1.33	1.93	0.84

FIG. 3 shows schematic illustrations of the arrangement according to the invention for different configurations, with the axially movable focus lens and an axially movable lens component for a correction mechanism:

a: with a cover glass and a long working distance,

b: with a cover glass and a short working distance,

c: without a cover glass and with a long working distance,

d: without a cover glass and with a short working distance.

The configurations with a long working distance refer to a focus shift into deep regions of the sample medium. The configurations with a short working distance have a focus position at a shallow sample depth. They only differ by the position of the focus lens. By an additional movement of the correction mechanism, further configurations with a cover glass and without a cover glass are possible. The correction mechanism is not moved for the focusing.

FIG. 4 illustrates transverse aberrations in μm for an axis point h′=0 mm and for a field edge point h′=9 mm, using light of the wavelengths 546 nm, 644 nm, 480 nm and 830 nm for the various configurations a, b, c, and d in FIG. 3. Continuous lines refer to the meridional section, and dashed lines to the sagittal section.

The individual aberration curves only differ to a very small degree. This shows that, according to the invention, neither the correction mechanism nor the focusing movement has a noticeable influence on the image quality.

FIG. 5 shows the arrangement according to the invention in a plan apochromat 25×/0.8 W for a focal range of 250 μm in an objective modification known from U.S. Pat. No. 7,782,539.

Proceeding from the object plane OE, the arrangement consists of a frontal group FG, having the lenses L1 and L2, an axially movable lens L3 designed as a focus lens FL, and a rear group RG with the lenses L4 to L11. The figure also shows the radii r1 to r17 of the individual optical components.

The schematic illustration of the arrangement according to the invention in FIG. 6 complements the illustration in FIG. 5 by adding the thicknesses D1 to D11 of the lenses L1 to L11, as well as the air spaces LA1 to LA7.

For a focal range of 250 μm, a numerical aperture of 0.8, a magnification factor β with a 25× larger image, and when an immersion agent/sodium chloride solution W is used, with a field number of 18, this embodiment is characterized by the following design data, wherein r1 to r17 are the curvature radii of the optical components in mm, D is the thickness of the air spaces between the optical components in mm, LA1 to LA7 are the air spaces between the optical components in mm, n_eis the refractive index, and v_eis the Abbe coefficient:


	Curvature		Refractive	Abbe
Lens	radius	Size	index	coefficient
No.	[mm]	[mm]	n_e	v_e

Sample, water	flat	P	1.33	55.8
Cover glass	flat	017	1.52	59.2
immersion	flat	060	1.33	55.8

1

flat

D1

0.50

1.49

70.2

2

r1

−1.654

D2

4.99

1.89

40.5

r2

−5.733

LA1

3

r3

−16.879

D3

1.50

1.52

64.0

r4

−7.899

LA2

4	r5	15.656	D4	2.00	1.59	68.0
	r6	34.321	LA3	1.65
5	r7	10.519	D5	4.50	1.55	45.5
	r8	9.906	LA4	2.30
6	r9	−43.509	D6	1.10	1.64	42.2
7	r10	9.045	D7	5.00	1.53	76.6
	r11	−13.679	LA5	0.10
8	r12	23.291	D8	5.80	1.53	76.6
9	r13	−7.743	D9	1.10	1.62	44.3
	r14	393.143	LA6	0.36
10	r15	10.029	D10	6.85	1.82	46.4
	r16	6.941	LA7	4.20
11	r17	−6.809	D11	2.83	1.60	35.0

r18	−7.687	0.15
		126.50

and a tube lens arranged behind the arrangement, the tube lens having a focal length of f=164.5 mm.

Depending on the sample thickness P, the cover glasses DE which are used, and the working distances AA, the following values are then found for the air spaces LA1 to LA2:


P	LA1	LA2
[mm]	[mm]	[mm]

0	1.42	1.19
0.25	0.50	2.11

The air spaces LA3 and LA7 are always the same in this case.

FIG. 7 shows schematic illustrations of the arrangement according to FIG. 5 for various focal depths in the sample, with a long working distance LA and a short working distance KA, and the axially movable focus lens FL.

FIG. 8 shows transverse aberrations in μm for an axis point h′=0 mm and for a field edge point h′=9 mm, using light of the wavelengths 546 nm, 644 nm, 480 nm and 830 nm for the various working distances LA and KA. Continuous lines refer to the meridional section, and dashed lines to the sagittal section.

Once again, the minimum changes in the aberration curves show the inventive function of the focusing arrangement at a constant image quality.

FIG. 9 shows the arrangement according to the invention in a plan apochromat 20×/1.0 W for a focal range of 300 μm in an objective modification known from DE 102011109783.

Proceeding from the object plane OE, the arrangement consists of a frontal group FG, having the lenses L1, L2, and L3, an axially movable lens L4 designed as a focus lens FL, and a rear group RG with the lenses L5 to L15. The figure also shows the radii r1 to r24 of the individual optical components.

The schematic illustration of the arrangement according to the invention in FIG. 10 complements the illustration in FIG. 9 by adding the thicknesses D1 to D15 of the lenses L1 to L15, as well as the air spaces LA1 to LA8.

For a focal range of 300 μm, a numerical aperture of 1.0, a magnification factor β with a 20× larger image, an immersion agent/sodium chloride solution W, and a field number of 20, this embodiment is characterized by the following design data, wherein r1 to r24 are the curvature radii of the optical components in mm, D is the thickness of the air spaces between the optical components in mm, LA1 to LA8 are the air spaces between the optical components in mm, n_eis the refractive index, and v_eis the Abbe coefficient:


	Curvature	Size	Refractive	Abbe
Lens No.	radius [mm]	[mm]	index n_e	coefficient v_e

Sample, water	flat	P	1.34	55.4
immersion	flat	2.62	1.34	55.4

1	r1	−11.117	D1	2.20	1.63	53.0
2	r2	−3.919	D2	1.41	1.76	52.1
3	r3	−21.906	D3	6.00	1.89	40.5

r4

−10.061

LA1

4

r5

−41.589

D4

4.00

1.73

28.3

r6

−24.293

LA2

5	r7	−104.081	D5	5.00	1.44	94.5
	r8	−21.109	LA3	0.10
6	r9	18.565	D6	9.65	1.44	94.5
	r10	−72.224	LA4	0.37
7	r11	60.934	D7	6.00	1.44	94.5
8	r12	−22.685	D8	1.50	1.75	35.1
9	r13	12.541	D9	6.79	1.44	94.5
	r14	−36.716	LA5	0.10
10	r15	12.363	D10	7.20	1.44	94.5
11	r16	−19.563	D11	1.10	1.64	42.4
	r17	20.125	LA6	0.10
12	r18	7.723	D12	4.50	1.82	46.4
	r19	5.002	LA7	6.00
13	r20	−6.689	D13	1.90	1.52	52.1
14	r2	19.342	D14	7.07	1.53	76.6
	r22	−57.991	LA8	0.45
15	r23	−455.522	D15	3.80	1.75	35.0

r24	−18.748	0.29
		107.00

and a tube lens is arranged behind the arrangement, the tube lens having a focal length of f=164.5 mm.


P [mm]	LA1 [mm]	LA2 [mm]

0	1.8	0
0.29	0	1.8

The air spaces LA3 and LA8 are always the same in this case.

FIG. 11 shows schematic illustrations of the arrangement in FIG. 9 for various focal depths in the sample, with a long working distance LA and a short working distance KA, and the axially movable focus lens FL.

FIG. 12 shows transverse aberrations in μm for an axis point h′=0 mm and for a field edge point h′=10 mm, using light of the wavelengths 546 nm, 644 nm, 480 nm and 830 nm for the various working distances LA and KA. Continuous lines refer to the meridional section, and dashed lines to the sagittal section.

The minimum changes in the transverse aberrations between the different working distances show, as above, the inventive function of the focusing arrangement.

Various embodiments of systems, devices, and methods have been described herein. These embodiments are given only by way of example and are not intended to limit the scope of the claimed inventions. It should be appreciated, moreover, that the various features of the embodiments that have been described may be combined in various ways to produce numerous additional embodiments. Moreover, while various materials, dimensions, shapes, configurations and locations, etc. have been described for use with disclosed embodiments, others besides those disclosed may be utilized without exceeding the scope of the claimed inventions.

Persons of ordinary skill in the relevant arts will recognize that the subject matter hereof may comprise fewer features than illustrated in any individual embodiment described above. The embodiments described herein are not meant to be an exhaustive presentation of the ways in which the various features of the subject matter hereof may be combined. Accordingly, the embodiments are not mutually exclusive combinations of features; rather, the various embodiments can comprise a combination of different individual features selected from different individual embodiments, as understood by persons of ordinary skill in the art. Moreover, elements described with respect to one embodiment can be implemented in other embodiments even when not described in such embodiments unless otherwise noted.

Although a dependent claim may refer in the claims to a specific combination with one or more other claims, other embodiments can also include a combination of the dependent claim with the subject matter of each other dependent claim or a combination of one or more features with other dependent or independent claims. Such combinations are proposed herein unless it is stated that a specific combination is not intended.

For purposes of interpreting the claims, it is expressly intended that the provisions of 35 U.S.C. §112(f) are not to be invoked unless the specific terms “means for” or “step for” are recited in a claim.

LIST OF REFERENCE NUMBERS

L1 to L15 lenses
r1 to r24 radii
D1 to D15 thicknesses
LA1 to LA8 air spaces
FL focus lens
FG frontal group
RG rear group
OE object plane
BE image plane
LA long working distance
KA short working distance
β magnification factor
P sample thickness
n_erefractive index
v_eAbbe coefficient

Claims

The invention claimed is:

1. An arrangement for changing the focus of a microscope objective comprising a plurality of optical components or component groups, wherein at least one of the optical components or component groups can move axially in an interior of the microscope objective, wherein the axially movable component or the axially movable component group includes a concave-convex lens or a lens group with a form factor according to the formula:

X = \frac{c_{1} + c_{2}}{c_{1} - c_{2}}

wherein X denotes the form factor, c₁is the curvature of a surface of the concave-convex lens or lens group facing an object plane, and c₂is the curvature of a surface of the concave-convex lens or lens group facing an image plane, and wherein X lies within a range from −8<X<−1, and wherein the concave-convex lens or lens group is arranged, proceeding from the object plane, behind a frontal group including first optical component or a component group, wherein a total focal length f_frontof the frontal group relative to a focal length f_objectiveof the objective lies in the range of:

f _front<−4×f _objectiveor f _front>4×f _objective.

2. The arrangement for changing the focus of a microscope objective according to claim 1, wherein, for an axial range of travel of the concave-convex lens or lens group between 0 mm and 5 mm, the focal length f is within the range from:

3×f _objective <f _ShiftLens<20×f _objective.

3. The arrangement for changing the focus of a microscope objective according to claim 1, wherein for a 25×/0.9 microscope objective, an immersion agent W including a sodium chloride solution, a focal range of 200 μm, a numerical aperture of 0.9, a magnification factor β with a 25× larger image, a field number of 18, and wherein r1 to r22 are the curvature radii of the optical components in mm, D is the thickness in mm, LA1 to LA6 are the air spaces between the optical components in mm, n_eis the refractive index, and v_eis the Abbe coefficient, the following design data characterizing the arrangement:

Size Curvature Air space D, Refractive Abbe Lens No. radius r [mm] LA [mm] index n_e coefficient v_e Sample, cell flat P 1.38 63.0 medium Cover glass flat DG 1.53 54.3 Immersion flat AA 1.34 55.4 1 flat D1 0.50 1.49 70.2 2 r1 −1.636 D2 5.79 1.89 40.5 r2 −6.167 LA1 3 r3 −14.549 D3 2.40 1.59 61.0 r4 −9.422 LA2 4 r5 70.312 D4 3.00 1.81 46.2 r6 −22.736 LA3 5 r7 42.712 D5 3.80 1.49 84.1 6 r8 −18.905 D6 1.60 1.66 39.5 7 r9 13.868 D7 3.90 1.50 81.1 r10 −18.518 LA4 8 r11 −344.203 D8 1.40 1.64 42.2 9 r12 8.092 D9 5.04 1.53 76.6 r13 −48.460 LA5 0.35 10 r14 8.015 D10 3.80 1.59 61.0 11 r15 11.802 D11 1.40 1.72 53.6 r16 5.902 LA6 5.14 12 r17 −6.704 D12 4.69 1.81 46.2 r18 −9.177 0.13 126.50

and a tube lens is arranged behind the arrangement, the tube lens having a focal length f of f=164 mm.

4. The arrangement for changing the focus of a microscope objective according to claim 3, wherein the design data include the following air spaces LA1 to LA4 in mm, according to different variations between sample thicknesses P in mm, cover glass thicknesses DG in mm, and immersions W with the thicknesses AA in mm:

P DG AA LA1 LA2 LA3 LA4 [mm] [mm] [mm] [mm] [mm] [mm] [mm] 0 0 0.56 1.78 0.49 2.47 0.3 0.2 0 0.56 0.95 1.33 2.47 0.3 0 0.17 0.4 1.78 0.49 1.93 0.84 0.2 0.17 0.4 0.95 1.33 1.93 0.84.

5. The arrangement for changing the focus of a microscope objective according to claim 1, wherein for a 25×/0.8 microscope objective, immersion agent W including a sodium chloride solution, a focal range of 250 μm, a numerical aperture of 0.8, a magnification factor β with a 25× larger image, a field number of 18, and wherein r1 to r17 are the curvature radii of the optical components in mm, D is the thickness in mm, LA1 to LA7 are the air spaces between the optical components in mm, n_eis the refractive index, and v_eis the Abbe coefficient, the following design data characterizing the arrangement:

Curvature Size Refractive Abbe Lens No. radius [mm] [mm] index n_e coefficient v_e Sample, flat P 1.33 55.8 water Cover glass flat 017 1.52 59.2 Immersion flat 060 1.33 55.8 1 flat D1 0.50 1.49 70.2 2 r1 −1.654 D2 4.99 1.89 40.5 r2 −5.733 LA1 3 r3 −16.879 D3 1.50 1.52 64.0 r4 −7.899 LA2 4 r5 15.656 D4 2.00 1.59 68.0 r6 34.321 LA3 1.65 5 r7 10.519 D5 4.50 1.55 45.5 r8 9.906 LA4 2.30 6 r9 −43.509 D6 1.10 1.64 42.2 7 r10 9.045 D7 5.00 1.53 76.6 r11 −13.679 LA5 0.10 8 r12 23.291 D8 5.80 1.53 76.6 9 r13 −7.743 D9 1.10 1.62 44.3 r14 393.143 LA6 0.36 10 r15 10.029 D10 6.85 1.82 46.4 r16 6.941 LA7 4.20 11 r17 −6.809 D11 2.83 1.60 35.0 r18 −7.687 0.15 126.50

and a tube lens arranged behind the arrangement, the tube lens having a focal length f of f=164 mm.

6. The arrangement for changing the focus of a microscope objective according to claim 5, wherein the design data include the following air spaces LA1 to LA2 in mm, according to various sample thicknesses P in mm:

P [mm] LA1 [mm] LA2 [mm] 0 1.42 1.19 0.25 0.50 2.11.

7. The arrangement for changing the focus of a microscope objective according to claim 1 wherein, for a 20×/1.0 microscope objective, immersion agent W including a sodium chloride solution, a focal range of 300 μm, a numerical aperture of 1.0, a magnification factor β with a 20× larger image, and a field number of 20, and wherein r1 to r24 are the curvature radii of the optical components in mm, D is the thickness in mm, LA1 to LA8 are the air spaces between the optical components in mm, n_eis the refractive index, and v_eis the Abbe coefficient, the following design data characterizing the arrangement:

Curvature Size Refractive Abbe Lens No. radius [mm] [mm] index n_e coefficient v_e Sample, flat P 1.34 55.4 water immersion flat 2.62 1.34 55.4 1 r1 −11.117 D1 2.20 1.63 53.0 2 r2 −3.919 D2 1.41 1.76 52.1 3 r3 −21.906 D3 6.00 1.89 40.5 r4 −10.061 LA1 4 r5 −41.589 D4 4.00 1.73 28.3 r6 −24.293 LA2 5 r7 −104.081 D5 5.00 1.44 94.5 r8 −21.109 LA3 0.10 6 r9 18.565 D6 9.65 1.44 94.5 r10 −72.224 LA4 0.37 7 r11 60.934 D7 6.00 1.44 94.5 8 r12 −22.685 D8 1.50 1.75 35.1 9 r13 12.541 D9 6.79 1.44 94.5 r14 −36.716 LA5 0.10 10 r15 12.363 D10 7.20 1.44 94.5 11 r16 −19.563 D11 1.10 1.64 42.2 r17 20.125 LA6 0.10 12 r18 7.723 D12 4.50 1.82 46.4 r19 5.002 LA7 6.00 13 r20 −6.689 D13 1.90 1.52 52.1 14 r2 9.342 D14 7.07 1.53 76.6 r22 −57.991 LA8 0.45 15 r23 −455.522 D15 3.80 1.75 35.0 r24 −18.748 0.29 107.00

and a tube lens is arranged behind the arrangement, the tube lens having a focal length f of f=164.5 mm.

8. The arrangement for changing the focus of a microscope objective according to claim 7, wherein the design data include the following air spaces LA1 to LA2 in mm, according to various sample thicknesses P in mm:

P [mm] LA1 [mm] LA2 [mm] 0 1.8 0 0.29 0 1.8.